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软体动物碳酸钙贝壳的形成过程涉及到Ca2+的吸收、转运、贮藏、分泌和沉积等,与钙代谢这一高度受控的复杂生理过程紧密相关。本研究以栉孔扇贝转录组测序结果为基础,通过RACE技术,从栉孔扇贝外套膜中克隆得到钙调素类似蛋白的cDNA序列,全长863bp,编码149个氨基酸,相应蛋白质的预测分子量约为17.0ku,理论等电点为4.03。钙调素类似蛋白具有4个可能的EF-hand Ca2+结合结构域,属于EF-hand钙结合蛋白家族,其氨基酸序列与栉孔扇贝钙调素钙调素的相似度为66%。半定量RT-PCR检测显示,钙调素类似蛋白基因在外套膜中特异性高表达,预示其可能参与贝壳的矿化过程。实时定量PCR检测显示,钙调素类似蛋白基因的表达水平随环境中Ca2+浓度的升高呈先升后降趋势,表明钙调素类似蛋白与外套膜中的钙代谢过程密切相关;在贝壳缺刻损伤后的再生过程中,钙调素类似蛋白基因表达量显著上升,暗示其积极参与到了贝壳的再生过程中。上述结果为进一步阐明钙调素类似蛋白基因的功能及其在栉孔扇贝生物矿化过程中的作用提供可能的理论依据。
Mollusc formation of calcium carbonate shells involves Ca2 + absorption, transport, storage, secretion and deposition, and calcium metabolism is closely related to the highly controlled complex physiological processes. In this study, based on the sequencing results of the scallop transcriptome of Chlamys farreri, the cDNA sequence of Calmodulin-like protein was cloned from the mantle of Chlamys farreri by RACE technology. The full-length cDNA was 863bp encoding a protein of 149 amino acids. The predicted molecular weight of the corresponding protein Is 17.0ku, the theoretical isoelectric point is 4.03. The calmodulin-like protein has four possible EF-hand Ca2 + binding domains and belongs to the EF-hand calcium binding protein family. The similarity of the amino acid sequence to calmodulin calmodulin is 66%. Semiquantitative RT-PCR showed that the calmodulin-like protein was highly expressed in the mantle, indicating that it may be involved in the mineralization of the shellfish. Real-time quantitative PCR showed that the expression levels of calmodulin-like protein genes increased first and then decreased with the increase of Ca2 + concentration in the environment, indicating that calmodulin-like proteins are closely related to calcium metabolism in the mantle; During regeneration, the expression of calmodulin-like protein gene increased significantly, suggesting that it is actively involved in the regeneration of the shellfish. The above results provide a possible theoretical basis for further elucidation of the function of calmodulin-like protein gene and its role in the biomineralization of Chlamys farreri.